Systems and devices for quantitative detections of biological data applications typically use a ticket, on which a target under test is placed. In the applications of drug discovery, medicine research and disease diagnostics, the detection targets include, but are not limited to, various cykotines such as Vascular Cell Adhesion Molecule-1 (VCAM-1), Interferon-γ (IFN-γ), Interleukin-6 (IL-6), and Interleukin-10 (IL-10) in human plasma, blood, urine and other body fluids. In the applications of bio-defense, the detection targets include, but are not limited to, various biological agents such as vaccinia, ricin, botulinum toxin and B. anthrax spores in water.
FIGS. 1A and 1B respectively illustrate top view and a side view of a lateral flow-based immunoassay ticket configuration. An adsorbent pad (a) receives a sample target and a conjugate release pad (b) includes a conjugate comprising of gold and antibody embedded therein. The sample passes through the conjugate release pad (b) and flows on a membrane (c) by a capillary flow. A zone (d) contains captured antibody (testing line), where antibody-antigen-antibody-gold complex (sandwich) is formed. A zone (e) contains control antibody where a control line is formed through direct antibody against another anti-species antibody. A receiving pad (f) receives liquid from the membrane (c).
FIG. 2 is an illustration of positive and negative immunoassay tickets. The assay includes four components: a capture antibody, an antigen, a detector antibody for binding the target, and a labeled reporter molecule of interest which binds to the detector antibody. The sample liquid is added into one or more sample well 22, also denoted as “S”. The control points or lines determine if the ticket itself is a functional ticket. In other words, if the control lines/points do not appear, the ticket is a bad ticket, regardless of the sample. For negative sample results, only control points or lines appear in the control zone 26, also denoted as “C”. For positive sample results, in addition to the control points or lines, there are target points or lines appearing in the target zone/area 24, also denoted as “T”. The ticket window area in FIG. 2 is the inner rectangle that includes the control zone/area and the target zone/area.
The reporter can be an enzyme, a fluorophore, a colored particle, a dyed particle, a particle containing a dye, a stained particle, a radioactive label, quantum dots, nanocrystals, up-converting phosphorescent particles, metal sols, fluorophore or dye containing polymer or latex beads that are detectable visually and/or with mechanical assistance and the like.
Because there are manufacturing variations among different ticket lots, the reader needs to be calibrated (compensated) for the manufacturing variations. The calibration needs to be in accordance with the specific ticket lot to be more effective
Therefore, there is a need for an accurate image-based biological data quantification device with ticket calibration capability.